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Hepatitis C virus detection related peptide and visible time-resolved fluorescent microsphere test strip thereof

A test strip and fluorescent technology, applied in the field of related peptides, can solve the problems of easy interference, no discovery, complex antibodies, etc., and achieve the effect of fast response, high accuracy and strong specificity

Active Publication Date: 2022-01-11
苏州华益美生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the development, and in the development of a large number of immunochromatographic test strip products before, the inventor has noticed that the key intermediate product neglected by people, that is, the quality control peptide, and the antibody produced by it cannot interfere with the detection reaction, but , the antibodies produced by HCV C, E1, E2, p7, NS2, NS3, NS4A, NS4B, NS5A and NS5B proteins and the antibodies produced by the human body due to infection with other pathogens are very complex and easily cause interference, resulting in the need for re-measurement
For this reason, the inventor carefully developed the quality control peptide without inspiration, and no errors caused by the quality control peptide have been found in the detection of actual samples so far.

Method used

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  • Hepatitis C virus detection related peptide and visible time-resolved fluorescent microsphere test strip thereof
  • Hepatitis C virus detection related peptide and visible time-resolved fluorescent microsphere test strip thereof
  • Hepatitis C virus detection related peptide and visible time-resolved fluorescent microsphere test strip thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 quality control peptide of the present invention

[0025] ①Preparation of quality control peptide-BSA (bovine serum albumin)

[0026] a) Quality control peptide sequence:

[0027] We have developed a quality control peptide (hapten) that does not interfere with the detection on our own platform, and synthesized the quality control peptide by synthetic method (synthetic unit: Sangon Bioengineering Shanghai (Co., Ltd.)). Its amino acid sequence is as shown in SEQ ID NO: 1.

[0028] b) Preparation of quality control peptide-BSA (i.e. a conjugate of quality control peptide and BSA)

[0029] Dissolve BSA with 0.05M MES pH6.0 to 10mg / ml, take 1ml and add 0.4mg of EDC, 1.1mg of NHS for activation for 30min, add 1mg of quality control peptide, couple for 2h, add 10μl of ethanolamine, block for 30 minutes, use 0.01 M PBS pH 7.4 for dialysis.

[0030] ②Preparation of quality control peptidomab

[0031] The quality control peptide-BSA was used as an antigen to im...

Embodiment 2

[0034] Example 2 Preparation of Hepatitis C Virus Core Antigen Monoclonal Antibody

[0035] According to the sequence of HCV cAg, we have developed two hapten fragments on our own platform and commissioned the synthesis of these two fragments (synthesis unit: Sangon Bioengineering Shanghai (Co., Ltd.)). The amino acid sequence of fragment 1 is shown as SEQ ID NO: 2, the amino acid sequence of Fragment 2 is shown in SEQ ID NO:3.

[0036] Basically refer to the method of step ①b) of Example 1, the difference is that the quality control peptide is replaced by the hepatitis C virus core antigen fragment 1 and 2 respectively, and the hepatitis C virus core antigen fragment 1-BSA and the hepatitis C virus core are respectively obtained Antigen fragment 2-BSA; basically refer to the method of step ② in Example 1, the difference is that the quality control peptide-BSA is replaced by hepatitis C virus core antigen fragment 1-BSA and hepatitis C virus core antigen fragment 2-BSA respect...

Embodiment 3

[0038] Example 3 Preparation of HCV antigen labeled with visible time-resolved fluorescent microspheres

[0039] Basically refer to the method of step ③ in Example 1, the difference is that the hepatitis C virus antigen is used instead of the quality control peptide-BSA, and the ratio is to add 20 μg of antigen per mg of visible fluorescent microspheres to obtain visible time-resolved fluorescent microsphere labeling Hepatitis C virus antigen, wherein the amino acid sequence of the hepatitis C virus antigen is SEQ ID NO:4.

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Abstract

The invention provides a quality control peptide, an HCV hapten and an HCV antigen for HCV detection, and a test strip based on the quality control peptide, the HCV hapten and the HCV antigen. The quality control peptide, the HCV hapten and the HCV antigen can be used for simultaneously detecting an HCV core antigen and an HCV antibody in a fluorescent or naked-eye manner.

Description

technical field [0001] The invention belongs to the field of immunology, and in particular relates to an immunochromatographic test strip for rapid detection of hepatitis C virus antigen antibody based on visible time-resolved fluorescent microspheres and related peptides therein. Background technique [0002] Hepatitis C (hepatitis C, HCV) is one of world infectious disease, according to the statistics of the World Health Organization, the global HCV infection rate has reached 3%, and the high chronicity is the maximum characteristic of this disease, the anti-HCV that produces can exist It lasts for several years to more than ten years in the patient's body, and even exists for life. Compared with hepatitis B, it is significantly more harmful than hepatitis B. 80% of patients with hepatitis C will develop chronic hepatitis, and a considerable proportion will develop into liver cirrhosis and liver cancer. , bringing long-term pain and huge economic burden to patients, their ...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K14/18G01N33/576G01N33/58G01N33/543G01N33/52
CPCC07K7/08C07K14/005G01N33/5767G01N33/582G01N33/54313G01N33/52C12N2770/24222G01N2333/186G01N2469/10G01N2469/20Y02A50/30
Inventor 张鑫王雪李晓李鹏昆杨晓娟尹立龙郭兰英李振勇
Owner 苏州华益美生物科技有限公司
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